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. 2019 Mar 6;9:3736. doi: 10.1038/s41598-019-40091-x

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Transcriptome analysis of ameloblast-like cells derived from gingival epithelial cells. (A) Validation of molecular differences among the fibroblasts and epithelial cells. Relative mRNA expressions of ameloblastic and osteogenic markers were analyzed by qPCR in different states of gingival cells. For differentiation, cells were treated with BMP4, BMP2, and TGFβ-1 for 7 days. a, expression of amelogenin; b, expression of ameloblastin; c, expression of osteopontin. 1 & 3, gene expression in cells without cytokines; 2 & 4, gene expression in cells treated with cytokines. Statistical significance of *P < 0.1, **P < 0.01, or ***P < 0.001 was determined by using Student t-test from three attempts performed by a limited culture number of 10 individual cultures. (B) The number of up- and down-regulated genes identified from the three comparison groups (GF vs dGE, dGF vs dGE, and GE vs dGE). Overlapping areas in Venn diagram represent common genes between the comparison groups. dGE/amelo, gingival epithelial cells treated with cytokines; GE, gingival epithelial cells without cytokines; dGF, gingival fibroblasts treated with cytokines; GF, gingival fibroblasts without cytokines. See also Table S2, Figs S1 and S2. (C) Heat map of 1007-differentially expressed genes (DEGs) from the pairwise comparison. Rows and columns represent 1007 DEGs (537 up- and 470 down-regulated genes) and profiled samples, respectively. The relative expression was depicted according to the color scale. See also Figs S3, S4, and S5. (D) Gene expression profile of the 20 genes selected by association with three major GO categories ‘cell surface’, ‘extracellular structure organization’, and ‘regulation of cell adhesion’. The heat map shows log₂FPKM values for 20 selected genes (rows) and four different states of cell (columns). Three GO categories were arranged by hierarchical clustering at left columns (cell surface in green, extracellular structure organization in purple, and regulation of cell adhesion in olive). (E) Confirmation of individual gene expression by qRT-PCR. Statistical significance of *P < 0.1, **P < 0.01, or ***P < 0.001 was determined by using Student t-test. GE, gingival epithelial cells without cytokines; dGE/amelo, gingival epithelial cells treated with cytokines.